Hydrophilic/Omniphobic Droplet Arrays for High-throughput and Quantitative Enzymology.

ABSTRACT: Engineered enzymes with enhanced or novel functions are specific catalysts with wide-ranging applications in industry and medicine. Here, we introduce droplet array microfluidic enzyme kinetics (DA-MEK), a high-throughput enzyme screening platform that combines water-in-air droplet microarrays formed on patterned superhydrophilic/omniphobic surfaces with cell-free protein synthesis to enable cost-effective expression and quantitative kinetic characterization of enzyme variants. By printing DNA templates encoding enzyme variants onto hydrophilic spots, stamping slides to add cell-free expression mix, and imaging the resulting arrays, we demonstrate reproducible expression of enzyme variants across hundreds of microwells per slide, with line of sight toward replicating this across larger lib

SUBMITTER: Lee B 

PROVIDER: S-EPMC12044592 | biostudies-literature | 2025 Apr

REPOSITORIES: biostudies-literature